PRODUCT CODE: ET1703-64

CD40L Recombinant Rabbit Monoclonal Antibody [JM11-34] (ET1703-64)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

-
+
Western blot analysis of CD40L on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1703-64, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: mouse liver tissue lysate<br />
Lane 2: rat liver tissue lysate<br />
Lane 2: mouse bone marrow tissue lysate
  • Western blot analysis of CD40L on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1703-64, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: mouse liver tissue lysate<br />
Lane 2: rat liver tissue lysate<br />
Lane 2: mouse bone marrow tissue lysate
  • ICC staining of CD40L in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-64, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of CD40L in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-64, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of CD40L in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-64, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CD40L antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-64, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of CD40L was done on THP-1 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-64, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of CD40L on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1703-64, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: mouse liver tissue lysate
Lane 2: rat liver tissue lysate
Lane 2: mouse bone marrow tissue lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

CD40L Recombinant Rabbit Monoclonal Antibody [JM11-34] (ET1703-64)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

Mouse liver tissue lysate, rat liver tissue lysate, mouse bone marrow tissue lysate, Hela, MCF-7, NIH/3T3, human spleen tissue, THP-1.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JM11-34

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

36 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

CD40L

SYNONYMS

CD 40L antibody; CD154 antibody; CD40 antigen ligand antibody; CD40 ligand antibody; CD40 ligand, soluble form antibody; CD40-L antibody; CD40L antibody; CD40L_HUMAN antibody; CD40LG antibody; gp39 antibody; hCD40L antibody; HIGM1 antibody; IGM antibody; IMD3 antibody; T B cell activating molecule antibody; T BAM antibody; T-cell antigen Gp39 antibody; TNF-related activation protein antibody; TNFSF5 antibody; TrAP antibody; Tumor necrosis factor (ligand) superfamily member 5 antibody; Tumor necrosis factor ligand superfamily member 5 antibody

SEQUENCE SIMILARITIES

Belongs to the tumor necrosis factor family.

TISSUE SPECIFICITY

Specifically expressed on activated CD4+ T-lymphocytes.

POST-TRANSLATIONAL MODIFICATION

The soluble form derives from the membrane form by proteolytic processing.; N-linked glycan is a mixture of high mannose and complex type. Glycan structure does not influence binding affinity to CD40.; Not O-glycosylated.

SUBCELLULAR LOCATION

Cell membrane, Cell surface, Secreted.

FUNCTION

Resting B cells can be activated and clonally expanded into antibody-producing cells in response to a combination of cell contact and soluble signals provided by primed helper T (Th) cells. While cytokines IL-4 and IL-13 alone are inadequate for B cell activation, contact with Th cells seems to be sufficient for delivery of proliferative signals. CD40 and CD154 (also designated CD40L or TRAP) comprise a receptor ligand pair central to the transmission of this signal. CD40 is expressed on the surface of B cells and CD154 is expressed on activated T cells. In the presence of such stimulus, IL-4 and IL-13 are capable of triggering immunoglobulin class switching and secretion of IgE. CD154 is a 261 amino acid protein that is is expressed as a soluble cytokine as well as a homotrimeric type II transmembrane protein. Its expression is tightly regulated, and abnormal levels of CD154 are associated with the pathogenesis of atheromatous plaque destabilization and thrombotic events. Mutations in the gene encoding for CD154 are implicated in hyper-IgM immunodeficiency syndrome type 1.